Dasgupta Shalini, Adhikari Jaideep, Das Priyanka, Sarkar Shubhanshu, Datta Pallab, Roy Chowdhury Amit, Barui Ananya
Center for Healthcare Science and Technology, Indian Institute of Engineering Science and Technology, Shibpur, Howrah 711103, India.
School of Advanced Materials, Green Energy and Sensor Systems, Indian Institute of Engineering Science and Technology, Shibpur, Howrah 711103, India.
ACS Appl Bio Mater. 2025 Jul 21;8(7):5644-5659. doi: 10.1021/acsabm.5c00278. Epub 2025 Jul 10.
Perfusable and functional vascularization is a critical yet unresolved challenge in regenerative medicine, particularly for large tissue constructs where passive diffusion of oxygen and nutrients is insufficient to sustain cellular viability. Without stable microvascular networks, engineered tissues often fail to survive or integrate following implantation, making vascularization a key determinant of clinical success. Conventional strategies─including growth factor supplementation, gene therapy, and endothelial coculture─have demonstrated limited efficacy due to inadequate vasculogenic signaling and compromised cell viability in the engineered niche. In this study, we report the development of an exosome-enriched biocomposite bioink composed of chitosan, collagen, and fibrinogen, incorporating vitamin D3 and periodontal ligament stem cell-derived exosomes (CCF-D3-exo). This formulation was used to fabricate perfusable three-dimensional (3D) vascular channel scaffolds via extrusion-based bioprinting, followed by the coculture of human adult dermal fibroblasts (HADF) and dental pulp stem cells (DPSC) within a dynamic lab-on-a-chip system. CCF-D3-exo scaffolds demonstrated significantly ( < 0.05) enhanced endothelial transdifferentiation and vasculogenic activity compared to exosome-free controls, with transcriptomic analysis showing upregulation of key angiogenic and vasculogenic markers, followed by proteomic validation via immunofluorescence, revealing robust expression and organized localization of α-SMA, vimentin, VEGF-A, VEGF-R1 CD34, and F-actin at different time points, indicative of early vascular morphogenesis and lumen formation. Together, these results establish a functional, perfusable, and biomimetic 3D vasculogenic niche, offering a promising strategy for fabricating vascularized skin grafts and advancing the translational potential of regenerative tissue constructs.
可灌注且具有功能的血管化是再生医学中一项关键但尚未解决的挑战，特别是对于大型组织构建物而言，氧气和营养物质的被动扩散不足以维持细胞活力。没有稳定的微血管网络，工程组织在植入后往往无法存活或整合，这使得血管化成为临床成功的关键决定因素。传统策略，包括生长因子补充、基因治疗和内皮细胞共培养，由于血管生成信号不足以及工程微环境中细胞活力受损，已显示出有限的疗效。在本研究中，我们报告了一种富含外泌体的生物复合生物墨水的开发，该生物墨水由壳聚糖、胶原蛋白和纤维蛋白原组成，并掺入了维生素D3和牙周膜干细胞衍生的外泌体（CCF-D3-exo）。该配方用于通过基于挤出的生物打印制造可灌注的三维（3D）血管通道支架，随后在动态芯片实验室系统中对成人人类皮肤成纤维细胞（HADF）和牙髓干细胞（DPSC）进行共培养。与无外泌体对照组相比，CCF-D3-exo支架显示出显著（<0.05）增强的内皮细胞转分化和血管生成活性，转录组分析显示关键血管生成和血管发生标志物上调，随后通过免疫荧光进行蛋白质组学验证，揭示了α-SMA、波形蛋白、VEGF-A、VEGF-R1、CD34和F-肌动蛋白在不同时间点的强烈表达和有组织的定位，表明早期血管形态发生和管腔形成。总之，这些结果建立了一个功能性、可灌注且仿生的3D血管生成微环境，为制造血管化皮肤移植物和推进再生组织构建物的转化潜力提供了一种有前景的策略。
